Namboothiri, H. R.; Hu, C. Y.

Precise regulation of gene expression in batch bacterial cultures is challenging because the underlying dynamics vary with cellular physiological state over time. Although cell-silicon systems enable rapid, real-time optogenetic control, disturbance rejection remains difficult in batch culture because the plant dynamics shift across growth phases, limiting the effectiveness of fixed-gain controllers designed under constant-growth assumptions. Here, we present a multiscale model-guided feedback control framework for disturbance rejection in batch E. coli cultures. Frequency-response analysis shows that the input-output dynamics of gene expression depend strongly on growth phase, revealing operating-point-dependent limits on the disturbance rejection performance of a fixed-gain PID controller. To address this limitation, we develop two growth-aware control strategies: a gain-scheduled PID (PID-GS) controller that adapts to cellular physiological state, and a gain-scheduled feedback-feedforward controller (PID-GS-FF) that further compensates for growth perturbations. We also introduce a controller evaluation framework that identifies three distinct operating regimes for targeted experimental validation. Together, these results show that accounting for growth-state-dependent dynamics is necessary for robust disturbance rejection in batch culture and provide a control-oriented framework for regulating living systems with shifting operating conditions.